I'm trying to synthesize 4 thio UTP, but I'm losing my final product (I think
at the precipitation step). I can see my 4 thio UDP peak shift on HPLC
directly after my reaction and after phenol extraction. My existing protocol
calls for me to bring the volume of my reaction upto 500ul with 300mM
NaOAcetate and 0.1% SDS and then precipitate with 1 vol. 1:1 ethanol/acetone
(by the way my initial reaction contains 20mM ATP and 10 mM 4 thio UDP in 100
ul). I'm not much of a biochemist and no details were given in the protocol
for the precipitation step eg. temp, time, spin speed or length of spin. If
anyone has any suggestions or if anyone has any 4 thio UTP as a positive
control it would be greatly appreciated.
Thank you
Sean Rollins
Rollins.24 at osu.edu